GSM7926115: Paneth cell, untreated, 1; Mus musculus; RNA-Seq
The proximal small intestine (10cm segment within a 2-12cm region from the gastroduodenal junction) from an adult (female 20-week old) Defa4 Cre :Rosa26 tdTomato/tdTomato mouse was removed, rinsed with ice cold PBS, opened longitudinally, and then scraped with a glass slide to remove villi prior to crypt elution using chelation buffer (PBS containing 2mM EDTA and 1mM DTT). To generate a single-cell suspension for flow cytometric analysis and sorting, isolated crypts were incubated with dispase and DNaseI, cell suspensions were filtered through a 40-μm cell strainer and then stained with rat anti-EpCAM–fluorescein isothiocyanate IgG2a antibody  and DAPI. Rat IgG2a–fluorescein isothiocyanate was used as a control. The MoFlo XDP100 cytometer with a 100-μm nozzle was used for single-cell FACS sorting EpCAM+ tdTomato - and EpCAM+ tdTomato + cell populations. FACS sorted tdTomato+ Defa4 Cre -expressing Paneth cells were loaded onto Chromium Next GEM Single Cell 3' Library and Gel Bead Kit V2 for complementary DNA library preparation.